2021年10月27日 星期三
芳樟叶片和枝的转录组分析及生物炭复合肥对芳樟醇合成调控基因表达特性的影响
Analysis on transcriptomes from leaf and branch of Cinnamomum camphora var. linaloolifera and effect of biochar compound fertilizer on expression characteristics of regulator genes related to linalool synthesis
2021年 第30卷 第4期 页码[31-40]    下载全文[1.3MB]  
摘要

以芳樟(Cinnamomum camphora var. linaloolifera Fujita)为研究材料,对施肥前及施肥后第1、第3和第5个月叶片和枝中芳樟醇相对含量进行了比较;利用高通量测序平台构建了施肥前后芳樟的转录组数据库并注释unigenes的功能,对芳樟醇合成途径进行预测,并分析了相关调控基因的表达特性变化。结果显示:施肥前后芳樟叶片的芳樟醇相对含量均高于枝,且施肥后叶片和枝中的芳樟醇相对含量均显著提高;施肥后第3个月叶片和枝中芳樟醇相对含量均最高,分别为89.89%和84.83%。从芳樟转录组中共获得72 560个unigenes,总长度96 543 844 bp,N50值为2 054 bp,GC含量为43.19%,其中,注释到7个数据库的unigenes共有52 613个。在NR数据库中,芳樟与荷花(Nelumbo nucifera Gaertn.)和博落回〔Macleaya cordata (Willd.) R. Br.〕的同源序列匹配度较高,占比分别为27.09%和14.91%。通过GO数据库,共有39 186个unigenes注释到生物过程、细胞成分和分子功能3大类24亚类中。在KEGG数据库中,共有40 698个unigenes注释到细胞过程、环境信息处理、遗传信息加工、代谢和生物系统5大类19亚类中;根据KEGG数据库的注释结果,推测芳樟醇主要通过MEP(甲基赤藓醇磷酸)途径合成前体物质,并在LIS(芳樟醇合酶)的作用下生成芳樟醇。通过KOG数据库对基因功能进行预测,共有40 096个unigenes注释到24个类别中。从芳樟叶片和枝的转录组中共鉴定出23个与芳樟醇合成相关的基因,且各基因的表达具有组织特异性;施肥后第3个月,叶片中有18个基因表达水平上调,枝中有15个基因表达水平上调,在叶片和枝中均上调表达的基因包括3个DXS基因、1个DXR基因、2个HDS基因、1个HDR基因、2个IDI基因、1个GPPS基因和3个LIS基因,说明这些基因可能参与芳樟醇合成的调控。综合分析结果表明:施用生物炭复合肥后,芳樟叶片和枝中芳樟醇相对含量均显著提高,与芳樟醇合成调控基因表达水平上调有一定关系。

Abstract

 Taking Cinnamomum camphora var. linaloolifera Fujita as research materials, relative contents of linalool in leaf and branch before fertilization and in the first, third, and fifth months after fertilization were compared; the transcriptome databases of C. camphora var. linaloolifera before and after fertilization were constructed by using high-throughput sequencing platform. and the functions of unigenes were annotated, the synthesis pathway of linalool was predicted, and the variation of expression characteristics of related regulator genes were analyzed. The results show that the relative contents of linalool in leaf of C. camphora var. linaloolifera before and after fertilization are all higher than those in branch, and the relative contents of linalool in leaf and branch after fertilization all increase significantly; the relative contents of linalool in leaf and branch in the third month after fertilization are the highest, which are 89.89% and 84.83%, respectively. There are 72 560 unigenes obtained from the transcriptome of C. camphora var. linaloolifera in total, the total length is 96 543 844 bp, the N50 value is 2 054 bp, and the GC content is 43.19%, in which, there are 52 613 unigenes annotated to 7 databases. In NR database, C. camphora var. linaloolifera has relatively high homologous sequence identity with Nelumbo nucifera Gaertn. and Macleaya cordata (Willd.) R. Br., and the percentages are 27.09% and 14.91%, respectively. In total, 39 186 unigenes are annotated to 24 subcategories of 3 categories namely biological process, cellular component, and molecular function via GO database. In KEGG database, there are 40 698 unigenes annotated to 19 subcategories of 5 categories containing cellular process, environment information processing, genetic information processing, metabolism, and organismal system; according to the annotation result of KEGG database, it is speculated that the precursor of linalool is mainly synthesized via MEP (methyl erythritol phosphate) pathway, and linalool is subsequently formed under the action of LIS (linalool synthetase). The gene functions were predicted through KOG database, and 40 096 unigenes are annotated to 24 categories. In total, 23 genes related to linalool synthesis are identified in the transcriptomes from leaf and branch of C. camphora var. linaloolifera, and the expression of each gene are tissue-specific; in the third month after fertilization, 18 genes in leaf are up-regulated, 15 genes in branch are up-regulated, and genes up-regulated in both leaf and branch contain 3 DXS genes, 1 DXR gene, 2 HDS genes, 1 HDR gene, 2 IDI genes, 1 GPPS gene, and 3 LIS genes, indicating these genes may be involved in the regulation of linalool synthesis. The comprehensive analysis result shows that after biochar compound fertilizer application, the relative contents of linalool in leaf and branch of C. camphora var. linaloolifera increase significantly, which may be associated with the up-regulation of regulator genes of linalool synthesis.

关键词芳樟; 芳樟醇; 转录组测序; 调控基因; 生物炭复合肥
Key wordsCinnamomum camphora var. linaloolifera Fujita; linalool; transcriptome sequencing; regulator gene; biochar compound fertilizer
作者向双a,b, 陈德强a,b, 孙维红a,b, 肖琳a,b, 邹双全a,b
所在单位福建农林大学: a. 林学院, b. 自然生物资源保育利用福建省高校工程研究中心, 福建 福州 350002
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基金项目中央财政林业科技推广示范项目(闽[2020]TG07号); 福建省林业科技项目(2021FKJ19)